Biotechnology
Journal
DOI 10.1002/biot.200600139 Biotechnol. J. 2006, 1, 1158–1162
1158 © 2006 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Basmati rice belongs to indica group of rice, which con-
stitute a small but special group that is considered the
best in cooking quality. However, basmati possesses low
tillering ability and exhibit yield losses in the same
ecosystem under similar cultural practices. Following a
transgenic approach, several potentially useful cloned
genes can now be introduced into basmati rice for incre-
mental improvement of the commercial cultivars. Howev-
er, indica rice varieties have generally been less respon-
sive to in vitro culture [1, 2]. Likewise, high frequency so-
matic embryogenesis with distinct embryo formation has
been limited [3]. In the present study an attempt was
made to improve somatic embryogenesis and the fre-
quency of green plant regeneration in indica type basmati
rice varieties by addition of cefotaxime in the callus in-
duction and proliferation medium. The antibiotic cefo-
taxime (Omnatax
TM
, C
16
H
16
N
5
O
7
S
2
Na) has been reported
to enhance the frequency of plant regeneration in wheat
[4], pearl millet [5] and sorghum [6]. This is perhaps the
first report describing the beneficial effect of cefotaxime
on somatic embryogenesis and subsequent plant regen-
eration in indica type basmati rice.
Callus cultures were established from mature seed
embryos of three indica type basmati rice varieties: Pusa
basmati 1, Basmati 370 and Basmati 386. Dehusked
healthy seeds after surface sterilization with HgCl
2
(0.1%)
for 10 min were rinsed three times with sterile water and
aseptically placed on Murashige and Skoog (MS) [7] medi-
um supplemented with 2,4-dichlorophenoxyacetic acid
(2,4-D; 2.5 mg/L), and 6-furfurylaminopurine (kinetin;
0.5 mg/L). Calli thus obtained were maintained and mul-
tiplied through periodic subculturing on two media based
on MS salts supplemented with 2,4-D (2.5 mg/l) and
kinetin (0.5 mg/L) with (treatment) or without (control)
100 mg/L cefotaxime. Cefotaxime, which is commonly
recommended for human injections, was dissolved in
sterile water and sterilized by passing through membrane
filter (0.2 μm), and was added into molten medium before
solidification. The medium without cefotaxime served as
control. The cultures were incubated at 25 ± 2°C at
70–80% humidity. Primary and secondary calli cultures
were kept in total dark.
After 2–3 weeks of incubation, calli showing nodular
regions were examined and selected under stereomicro-
Short Communication
Influence of antibiotic cefotaxime on somatic embryogenesis
and plant regeneration in indica rice
Deepinder Grewal, Raman Gill and Satbir Singh Gosal
Department of Plant Breeding, Genetics and Biotechnology, Punjab Agricultural University, Ludhiana, India
An antibiotic, cefotaxime (Omnatax
TM
) has been found to promote somatic embryogenesis and
subsequent plant regeneration in vitro in indica-type basmati rice cultures. Response was highly
genotype specific. The number, mass and morphology of the calli formed on the scutellar tissues
were dependent on the growth medium (with or without cefotaxime). The embryogenic nature of
nodular calli was confirmed through histological analysis and their plant regeneration ability. The
calli of variety Pusa basmati 1 grown on medium supplemented with cefotaxime (100 mg/L) ex-
hibited up to 70.5% plant regeneration as compared to control (51.51%). Plants regenerated from
emryogenic calli were phenotypically normal and identical to seed-derived plants and exhibited
normal fertility. A limited humidity and an optimal aeration of the culture tubes further enhanced
the frequency of somatic embryogenesis and plant regeneration.
Keywords: Basmati rice · Cefotaxime · Oryza sativa · Shoot proliferation · Somatic embryogenesis
Correspondence: Dr. Deepinder Grewal, Department of Plant Breeding,
Genetics and Biotechnology, Punjab Agricultural University,
Ludhiana 141 004, India
E-mail: grewaldeepi@yahoo.co.in
Abbreviations: BAP, benzylaminopurine; 2,4-D, 2,4-dichlorophenoxyacetic
acid; kinetin, 6-furfurylaminopurine; MS, Murashige and Skoog
Received 31 July 2006
Accepted 07 September 2006